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Remedy for spinal injury containing interleukin-6 antagonist

a technology of interleukin-6 and spinal cord injury, which is applied in the direction of antibody medical ingredients, drug compositions, peptide/protein ingredients, etc., can solve the problems of side effects, no effective treatment methods are currently available, and no results that indicate recovery from paralysis, etc., and achieve the effect of restoring motion coordination and spinal cord recovery

Inactive Publication Date: 2006-07-27
CHUGAI PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0033] Thus, there is a need for a therapeutic means that not only prevents the aggravation of conditions of a spinal cord injury but also aids recovery therefrom, and the present invention provides a pharmaceutical composition useful as one of the means.
[0034] After intensive and extensive studies to resolve the above problem, the present inventors have found that agonist against IL-6, for example, antibody against IL-6 receptor has an effect of aiding recovery from spinal cord injury. Thus, the present invention provides a therapeutic agent for spinal cord injury comprising interleukin-6 (IL-6) antagonist as an active ingredient.

Problems solved by technology

As therapeutic regimens for spinal cord injury, no effective methods of treatment are currently available and there are only symptomatic treatments including local stabilization such as surgery.
In order to prevent the aggravation of pathological conditions, steroids have been administered in large quantities, but no results have been obtained that indicate recovery from paralysis.
It has been reported, however, that this method is associated with side effects such as the aggravation of diabetes mellitus and pneumonia.
However, the clinical application of the implantation of the fetus spinal cord is very difficult due to the shortage of donors and to ethical problems.
In fact, however, they do not differentiate into neurons after injury, and all differentiate into glia cells thereby forming scars.
However, there is no literature that demonstrates that spinal cord injury can be repaired by the differentiation induction of neural stem cells by cytokines.

Method used

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  • Remedy for spinal injury containing interleukin-6 antagonist
  • Remedy for spinal injury containing interleukin-6 antagonist
  • Remedy for spinal injury containing interleukin-6 antagonist

Examples

Experimental program
Comparison scheme
Effect test

example 1

Materials and Methods

Animals:

[0137] Adult (18-22 g) female c57BL / 6J mice were used in all experiment groups.

Spinal Cord Injury:

[0138] Female mice were anesthetized by the intraperitoneal injection of ketamine (100 mg / kg) and xylazine (10 mg / kg). The back was shaved, a 20 mm midline skin incision was made, and then the spine was exposed. After the thoracic region of the spine was exposed by separation to the sides of the back muscle, the spinous process of the T7-T13 vertebra was exposed. Laminectomy was made at the ninth thoracic spine level to expose the spinal cord taking care not to injure the dura mater. The spine was stabilized with forceps and clamps on the T7 and T11 spinous processes and the ligament. After the animal body was floated by lowering the stage, spinal cord injury (SCI) was made by the NYU impactor. A 3 g weight (the apex with a diameter of 1.2 mm) was dropped from a height of 25 mm to the T9 level spinal cord. Muscles and the incised part were closed in l...

reference example 1

Preparation of Human Soluble IL-6 Receptor

[0153] Soluble IL-6 receptor was prepared by the PCR method using a plasmid pBSF2R.236 containing cDNA that encodes IL-6 receptor obtained according to the method of Yamasaki et al., (Yamasaki, K. et al., Science (1988) 241, 825-828). Plasmid pBSF2R.236 was digested with a restriction enzyme Sph I to obtain the cDNA of IL-6 receptor, which was then inserted into mp18 (manufactured by Amersham). Using a synthetic oligoprimer designed to introduce a stop codon into the cDNA of IL-6 receptor, a mutation was introduced into the cDNA of IL-6 receptor by the PCR method using the in vitro Mutagenesis System (manufactured by Amersham). The procedure resulted in the introduction of a stop codon to the amino acid at position 345, and gave cDNA encoding soluble IL-6 receptor.

[0154] In order to express the cDNA of soluble IL-6 receptor in CHO cells, it was ligated to a plasmid pSV (manufactured by Pharmacia) to obtain a plasmid pSVL344. The cDNA of so...

reference example 2

Preparation of Anti-Human IL-6

[0157] antibody

[0158] Ten μg of the recombinant IL-6 (Hirano et al., Immunol. Lett., (1988) 17, 41) was used for immunizing BALB / c mice together with Freund's complete adjuvant, and this was repeated every week until anti-IL-6 antibody could be detected in the serum. Immune cells were extracted from local lymph nodes and were then fused with a myeloma cell line P3U1 using polyethylene glycol 1500. Hybridomas were selected according to the method of Oi et al. (Selective Methods in Cellular Immunology, W.H. Freeman and Co., San Francisco, 351, 1980) that employs the HAT medium, and the hybridoma that produces anti-human IL-6 antibody was established.

[0159] The hybridoma that produces anti-human IL-6 antibody was subjected to the IL-6 binding assay as follows. Thus, a 96-well microtiter plate made of flexible polyvinyl (manufactured by Dynatech Laboratories, Inc., Alexandria, Va.) was coated with 100 μl of goat anti-mouse Ig (10 μl / ml, manufactured by C...

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Abstract

A therapeutic agent for spinal cord injury, a modulator of differentiation of neural stem cells and an inhibitor of differentiation into glia cells comprising an interleukin-6 antagonist as an active ingredient.

Description

TECHNICAL FIELD [0001] The present invention relates to a therapeutic agent for spinal cord injury comprising interleukin-6 (IL-6) antagonist as an active ingredient. BACKGROUND ART [0002] In today's society, a person may suffer spinal cord injury due to a motor vehicle accident, a fall, a tumble, a sports injury and the like. In Japan, the annual number of the injured people is about 5,000 with a cumulative number of patients possibly amounting to 100,000. Symptoms of spinal cord injury are very severe including permanent quadriplegia, motor paralysis and sensory paralysis, bladder and rectum disorders, respiratory disorders etc., and the daily management thereof includes rehabilitation, respiratory management, bedsore prevention, the management of defecation and urination, and the like. [0003] As therapeutic regimens for spinal cord injury, no effective methods of treatment are currently available and there are only symptomatic treatments including local stabilization such as surg...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K38/00A61P19/00A61P25/00A61P43/00C07K16/24
CPCA61K38/00A61K2039/505C07K16/248C07K16/2866A61P19/00A61P25/00A61P43/00A61K39/395A61K45/00
Inventor OKANO, HIDEYUKIOKADA, SEIJINAKAMURA, MASAYAYOSHIZAKI, KAZUYUKI
Owner CHUGAI PHARMA CO LTD
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